Seasonal Variation of Proteins and Amino Acids Content in Various Parts of Aegle Marmelos (L.) Corr

Materials and Methods

1) The protein was quantitatively estimated by the Lowey, et al. [6] method. Chemicals: 0.1% N NaOH - (4gm in 1000ml), 2% Na2CO3 - (2gm in 100ml distilled water), 0.5% CuSO4- (0.5gm in 100ml distilled water), 1 % Na-K-tartarate and 5 % Trichloro acetic acid/per chloric acid. Reagents: Lowry A-2% Na2CO3in 0.1% N NaOH, Lowry B- 5% CuSO4 in 1% Na-K-tartarate and Lowry C-98ml A and 2 ml B, Lowry D - Folin phenol reagent. Procedure: 1gm of plant material was homogenized with 10ml, 80% ethanol. The extract was centrifuged at 5000 rpm for 5 min. and the supernatant was discarded. 5%, 10 ml Trichloro acetic acid (TCA) or Per chloric acid (PCA) was add to residue and incubated at 800C for 20 minutes. The pellet was centrifuged and the supernatant was discarded. Residue was washed with 10 ml distilled water and again recentrifuged. The supernatant was discarded. 2%, 10ml Na2CO3 in 0.1 N NaOH was added to the residue and incubated for an hour at 30°C and again centrifuged and residue was discarded. The final volume of supernatant was measured and it was used as a sample for protein. 1ml of aliquot of sample was taken and 5ml reagent C was added to it mixed it thoroughly. The sample was incubated for 10 minutes and 1ml of reagent D was added to it. The colour intensity was read at 660 nm. Using Spectrophotometer. The protein concentration of an unknown sample was calculated byusing standard graph.

The amino acids content of leaves was 0.613 mg/g dry wt. in summer, 0.542 mg/g dry wt. in winter and 0.423 mg/g dry wt. in monsoon. Higher being observed during summer i.e. 0.613 mg/g dry wt. The range of amino acids content in bark 0.388 mg/g dry wt. to 0.506 mg/g dry wt. Maximum concentration of amino acids was noted during summer 0.506 mg/g dry wt. The range of amino acid content of wood from 0.259 mg/g dry wt. to 0.404 mg/g dry wt. The amino acids content of root was lower (0.257 mg/g dry wt. to 0.330 mg/g dry wt.) as compared to leaves, stem and wood of all seasons. Generally, the concentration of amino acids were found to be in increasing order of root< wood< bark < leaves (Table 1 & Graph 1).

2) The estimation of total amino acid was carried out by Krishnamurthy, et al. [7] method. Reagents: Alcoholic ninhydrin. (100 ml alcohol + 400 mg ninhydrin) and Glycine (Std.) (10mg glycine + 100ml distilled water) Procedure: 500 mg plant material was ground in mortar and pestle with few drops of cold 80% ethanol. Then 2.5ml of distilled water and 10ml of boiling 80% ethanol were added to it. The extract was centrifuged for 15 minutes at 10,000rpm. Residue was discarded the supernatant was collected and total volume was made 15ml with distilled water. Test tube was kept at 60°C for 20 minutes. The test tube was cooled and 1ml 50% ethanol was added. Read at 420 nm in spectrometer. Glycine was used as stand rand.

Results and Discussion

The protein content of leaves was higher (5.964 mg/g dry wt.) in summer over than winter (5.546 mg/g dry wt.) and monsoon (4.264 mg/g dry wt.). The range of protein content of bark was noted from (5.647 mg/g dry wt. to 6.194 mg/g dry wt.). The ranges of protein content in wood were from (3.196 mg/g dry wt. to 4.435 mg/g dry wt.) and show higher in summer. The protein content of root was higher (4.295 mg/g dry wt.) in summer over than winter (3.529 mg/g dry wt.) and monsoon (3.288 mg/g dry wt.). The protein content of seeds was higher (26.648 mg/g dry wt.) as compared to leaves, stem and roots of all seasons. The protein content showed increasing order of root < wood <leaves<bark (Table 1 & Graph 1).

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